7. Project Summary/Abstract The developmental origins of childhood physical, emotional, and behavioral problems begin in utero. Prenatal experiences ?program? the infant for the context s/he will encounter upon birth, laying a foundation for health outcomes and disease. However, the biological mechanisms linking the prenatal environment to early infant outcomes remain understudied. The application of epigenetic methods to human behavior is a relatively new and innovative endeavor devoted to understanding how environmental influences shape gene expression independent of DNA structure. Infants exposed to extreme stress during pregnancy show epigenetic adaptations, consistent with theories that biological systems calibrate in preparation for a high-risk postnatal environment. Innovative human research on epigenetic processes is sorely needed. There are two major gaps in the literature to date. First, the mechanisms linking maternal stress to epigenetic outcomes are still poorly understood. Second, most epigenetic studies only examine a small number of marks on select (i.e., fewer than 10) candidate genes, rather than a complex interdependent genetic network. The objective of this proposal is to advance the science and technology of prenatal programing research by (1) identifying mothers with the full range of emotional distress and carefully characterizing maternal stress reactivity (e.g., autonomic and neuroendocrine) in a laboratory assessment; and (2) developing a novel, hypothesis-driven assay to assess epigenetic processes within a network of genes. These genes were selected based on rigorous review of published associations with HPA axis functioning. Consistent with the Research Domain Criteria (RDoC) framework, mothers will be screened and enrolled based upon a range of scores on emotion dysregulation (ED) measures. ED is an important transdiagnostic vulnerability that has been linked to nearly every psychiatric diagnosis, high stress exposure, parenting difficulties, health-risk behaviors, and intergenerational transmission of psychopathology. Specifically, we will recruit 158 mothers with a uniform distribution on ED measures, conduct a thorough laboratory assessment of stress, measure epigenetic outcomes in placental samples, assess newborn behavioral outcomes, and develop an assay of stress-related epigenetic markers. This innovative project is both comprehensive and economical; it unites a stellar team of scientists with equipment, many of the necessary supplies, and generous institutional support to conduct the proposed research. When the aims of this project are realized we will have an improved understanding of early outcomes for infants of dysregulated mothers. We will also have created a novel assay, which will promote rapid replication as well as new investigations of stress-related epigenetic marks. This short-term longitudinal study lays the foundation for further follow-up of infants into early childhood and a programmatic line of research devoted to understanding and intervening with dysregulated mothers and children, especially those in highest need of improved preventative care.